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浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2012, Vol. 38 Issue (6): 725-713    DOI: 10.3785/j.issn.1008-9209.2012.01.171
食品科学     
响应面分析法优化桑叶叶绿素提取工艺
陈绍瑗， 吕贞儿， 董峰丽， 毛碧增
浙江大学 农业与生物技术学院，浙江 杭州 310058
Optimization of chlorophyll extraction from mulberry leaves using response surface methodology
CHEN Shao-yuan, LÜ Zhen-er, DONG Feng-li, MAO Bi-zeng
College of Agriculture & Biotechnology, Zhejiang University, Hangzhou 310058, China
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摘要： 为优化桑叶叶绿素提取工艺，根据单因素试验结果与响应面分析法中的Box-Behnken中心组合设计原则选取试验因素与水平，从中选取对叶绿素提取结果有明显影响的提取时间、提取温度、提取液料比3个因素进行优化；并利用Design Expert 7.1.6分析软件对试验数据进行分析. 结果表明：桑叶叶绿素提取的最佳工艺参数为提取时间5.25 h，提取温度56.5 °C，液料比103∶1；在此条件下，桑叶叶绿素质量分数为5.376 mg/g，与预测值5.451 mg/g接近. 说明根据Box-Behnken模型、采用响应面分析法得到的桑叶叶绿素提取优化工艺准确可靠.
Abstract:   Chlorophyll plays an important role in our everyday life. It is used not only as an annexing agent in medicinal and calleidic products but also as an unartificial food coloring agent. Chlorophyll and its ramifications have antioxidant and anti-mutation properties. China has abundant mulberry resources. The mulberry leaf is an excellent Chinese herb from ancient times to the present day and contains many kinds of bioactive components, in which the chlorophyll content is an important physiological index. However, the optimization of chlorophyll extraction from mulberry leaf by response surface methodology (RSM) has not been reported so far. Response surface methodology (RSM) is an effectively statistical technique for optimizing complex processes. The greatest advantage of RSM is that we need fewer experimental trials. It is widely used in optimizing variables of the extraction process, such as polysaccharides, alkaloids, phenolic compounds and protein from various materials. Box-Behnken design (BBD) is one of RSM, which only has three-levels (low, medium and high, coded as −1, 0 and +1), and needs fewer experiments. It’s more efficient and easier to arrange and interpret experiments compared with others. This study focused on the optimal conditions of extracting mulberry leaf chlorophyll using RSM. The mixture of acetone and ethanol was used as extraction solvent. The chlorophyll content was calculated by Arnon’s method. Extraction time, temperature, ratio of liquor to material and volume ratio of acetone to ethanol in extraction buffer were selected as the experiment factors according to the results of single factor experiments. The extracting parameters were optimized by RSM. Under the optimal extraction conditions described above, the confirmatory experiment was conducted. The single factor experiment results indicated that extraction time of 5 h (Fig. 2) , extraction temperature of 50 °C (Fig. 3), ratio of liquor to material of 100∶1 (Fig. 4) and volume ratio of acetone to ethanol of 3∶1 (Fig. 1) were adopted for following experiment. Three independent variables were extraction time (C), extraction temperature (B) and ratio of liquor to material (A), according to the influence∶1. Under the optimal extraction condition, the chlorophyll productivity was 5.376 mg/g, which was in consistent with the predicted value of 5.451 mg/g.s of the four factors on the chlorophyll productivity. The statistical analysis indicated that the three variables (Table 4) and the quadratic of extraction time (C) and ratio of liquor to material (A) (Fig. 6) had significant effects on the yields, and there had the significant interaction effects between the variables of extraction temperature (B) and extraction time (C) (Fig. 7). A high determination co-efficient mathematical model was gained that could be used to optimize chlorophyll extraction. Design Expert 7.1.6 software showed the optimal extraction condition was as follows: extraction time of 5.25 h, extraction temperature of 56.5 °C, ratio of liquor to material of 103 In conclusion, the optimization of chlorophyll extraction by RSM is convenient, feasible and highly efficient
出版日期: 2012-11-20
基金资助:

浙江省科技厅公益性应用研究资助项目 (2011C22027)
通讯作者: 毛碧增，Tel: +86-571-88982678; E-mail: maobz@zju.edu.cn   
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引用本文:

陈绍瑗， 吕贞儿， 董峰丽， 毛碧增. 响应面分析法优化桑叶叶绿素提取工艺[J]. 浙江大学学报(农业与生命科学版), 2012, 38(6): 725-713.

CHEN Shao-yuan, Lü Zhen-er, DONG Feng-li, MAO Bi-zeng. Optimization of chlorophyll extraction from mulberry leaves using response surface methodology. , 2012, 38(6): 725-713.

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http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2012.01.171    或    http://www.zjujournals.com/agr/CN/Y2012/V38/I6/725

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